靶向c-myc基因的siRNA基因干扰对大肠癌Lovo细胞增殖与凋亡的影响

The effect of the interference on the si RNA with targeted c- myc gene on the proliferation and apoptosis of colorectal cancer Lovo cell

目的阐述靶向c-myc基因的si RNA基因转染干扰对大肠癌Lovo细胞增殖与凋亡的影响作用。方法将靶向c-myc基因的si RNA片段转染入大肠癌Lovo细胞,特异沉默c-myc基因,通过实时荧光定量PCR法检测c-myc m RNA表达水平,原位末端标记法(terminal deoxynucleotidyl transferase d UTP nick end labeling,TUNEL)检测肿瘤细胞凋亡情况及噻唑蓝法[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]检测细胞的增殖能力。结果经过转染的Lovo细胞组与未转染的对照组c-myc m RNA表达量比率为0.22∶1,c-myc m RNA表达量比例有统计学差异(P<0.05)。经过转染的Lovo细胞与未转染的对照组的凋亡指数分别为32.4%与65.4%,具有显著性统计学差异(P<0.01)。MTT检测经过转染的Lovo细胞与未转染的对照组的细胞增值率,分别为79.28%与38.32%,具有极其显著性统计学差异(P<0.001)。结论将靶向c-myc基因的si RNA片段转染入大肠癌Lovo细胞,特异沉默c-myc基因,从而证实c-myc抑制大肠癌细胞增殖,诱导细胞凋亡的作用,从而为大肠癌的基因治疗寻求更多的治疗靶点。

Objective To study the effects that interference of the si RNA with targeted c-myc gene has on the proliferation and apoptosis of colorectal cancer Lovo cell. Methods The si RNA clips with targeted c-myc gene was transfected into the colorectal cancer Lovo cells, then the specific c-myc gene was specifically silenced. The c-myc m RNA expression level was detected using realtime-fluorescence quantitative polymerase chain reaction（RT-PCR）, and the apoptosis of the tumor cell was detected through the terminal deoxynucleotidyl transferase d UTP nick end labeling（TUNEL） method, while the proliferation of the tumor cell was detected through the 3-（4, 5-dimethyl-2-thiazolyl）-2, 5-diphenyl-2-H-tetrazolium bromide（MTT）method. Results The rate of the expression of the c-myc m RNA of the transfected Lovo cell group and nontransfected group was 0.22：1, which was statistically significant（P〈0. 05）. The indices of apoptosis of the transfected Lovo cell group and non-transfected group were 32.4% and 65.4%, respectively, which was also statistically significan（P〈0. 01）. The rates of the cell proliferation detected by MTT of the transfected Lovo cellgroup and non- transfected group were 79.28% and 38.32%, respectively, which was extremely statistically significant（P〈0.001）. Conclusion After being targeted with the si RNA, and transfected into the colorectal cancer Lovo cells and silenced specifically, the c-myc gene inhibits the proliferation and induces the apoptosis of colorectal cancer cells, and could provide more therapeutic targets for gene therapy of colorectal cancer.